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Q1 – What type of hydrogel are TruSphere Inserts made with?
The insert are made of natural polysaccharide-based hydrogel.
Q2 – Is the hydrogel Xeno-free?
Yes. TruSphere Inserts contain no non-human biomolecules.
Q3 – Can TruSphere Inserts be embedded in paraffin?
Yes. The hydrogel structure of the insert can be directly embedded in paraffin blocks for tissue sectioning, allowing for spheroid and organoid slices to be obtained.
Q4 – Are TruSphere Inserts compatible with transwell assays?
Yes. The TruSphere 24-well inserts are compatible with transwell inserts, facilitating co-culture or immune cell infiltration studies.
Q5 – Are TruSphere Inserts compatible with ECM assays?
Yes. Extracellular matrix (ECM) components can be used to ECM embed formed spheroids/organoids in the microwells to create a comprehensive 3D tissue model.
Q6 – Can you seed microwells with a single cell?
Yes. Diluting a primary cell suspension will all a single cell or near-single cell in each of the microwells. Growing spheroids from these single cells depends on several factors, including cell type, phenotype, and the presence or absence of ECM. We recommend using a 24-well plate for this purpose. Single cells that are isolated in the microwells can be monitored and images captured through a bright-field microscope.
Q7 – How long can organoids be maintained in a TruSphere insert?
Spheroids and organoids may be maintained in a TruSphere microwell for weeks and months as long as no contamination issues arise.
Q8 – How would I perform a qPCR using TruSphere inserts?
For gene expression analysis, the spheroids/organoids should be removed from the plate, which can be easily accomplished by rinsing the inserts with the appropriate culture media before following a standard RNA extraction protocol.
Q9 – How would I perform a ICC assays using TruSphere inserts?
ICC, or any immunofluorescent analysis, can be performed without removing the organoids from the insert. Fixing, staining, and imaging can all be carried out while the organoids or spheroids remain in the microwells.
Q10 – How would I perform an IHC assay using TruSphere inserts?
Spheroids within the TruSphere inserts can use the same process for immunohistochemistry (IHC) analysis without needing to remove the organoids for paraffin embedding.